scholarly journals Antisense Oligonucleotide Screening to Optimize the Rescue of the Splicing Defect Caused by the Recurrent Deep-Intronic ABCA4 Variant c.4539+2001G>A in Stargardt Disease

Genes ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 452 ◽  
Author(s):  
Alejandro Garanto ◽  
Lonneke Duijkers ◽  
Tomasz Z. Tomkiewicz ◽  
Rob W. J. Collin
Keyword(s):  
Pluripotent Stem Cells ◽  
Antisense Oligonucleotides ◽  
Autosomal Recessive ◽  
Stargardt Disease ◽  
Single Nucleotide ◽  
Significant Correction ◽  
Nucleotide Mismatch ◽  
Single Nucleotide Mismatch ◽  
Splicing Defects ◽  
Induced Pluripotent

Deep-sequencing of the ABCA4 locus has revealed that ~10% of autosomal recessive Stargardt disease (STGD1) cases are caused by deep-intronic mutations. One of the most recurrent deep-intronic variants in the Belgian and Dutch STGD1 population is the c.4539+2001G>A mutation. This variant introduces a 345-nt pseudoexon to the ABCA4 mRNA transcript in a retina-specific manner. Antisense oligonucleotides (AONs) are short sequences of RNA that can modulate splicing. In this work, we designed 26 different AONs to perform a thorough screening to identify the most effective AONs to correct splicing defects associated with c.4539+2001G>A. All AONs were tested in patient-derived induced pluripotent stem cells (iPSCs) that were differentiated to photoreceptor precursor cells (PPCs). AON efficacy was assessed through RNA analysis and was based on correction efficacy, and AONs were grouped and their properties assessed. We (a) identified nine AONs with significant correction efficacies (>50%), (b) confirmed that a single nucleotide mismatch was sufficient to significantly decrease AON efficacy, and (c) found potential correlations between efficacy and some of the parameters analyzed. Overall, our results show that AON-based splicing modulation holds great potential for treating Stargardt disease caused by splicing defects in ABCA4.

scholarly journals The presence of a single-nucleotide mismatch in linker increases the fluorescence of guanine-enhanced DNA-templated Ag nanoclusters and their application for highly sensitive detection of cyanide

RSC Advances ◽  
2018 ◽  
Vol 8 (72) ◽  
pp. 41464-41471 ◽  
Author(s):  
Jun Peng ◽  
Jian Ling ◽  
Qiu-Lin Wen ◽  
Yu Li ◽  
Qiu-E. Cao ◽  
...  
Keyword(s):  
Sensitive Detection ◽  
Single Nucleotide ◽  
Single Base ◽  
Highly Sensitive ◽  
Nucleotide Mismatch ◽  
Single Nucleotide Mismatch ◽  
Ag Nanoclusters ◽  
Ag Ncs ◽  
Highly Sensitive Detection

Single-base mismatched G-rich enhanced DNA-Ag NCs for cyanide detection.

Switchable lanthanide luminescent binary probes in efficient single nucleotide mismatch discrimination

2015 ◽  
Vol 211 ◽  
pp. 297-302 ◽  
Author(s):  
Ulla Karhunen ◽  
Eeva Malmi ◽  
Ernesto Brunet ◽  
Juan Carlos Rodríguez-Ubis ◽  
Tero Soukka
Keyword(s):  
Single Nucleotide ◽  
Nucleotide Mismatch ◽  
Single Nucleotide Mismatch ◽  
Mismatch Discrimination

scholarly journals Integration-free induced pluripotent stem cells derived from a patient with autosomal recessive Alport syndrome (ARAS)

2017 ◽  
Vol 25 ◽  
pp. 1-5 ◽  
Author(s):  
Bernd Kuebler ◽  
Begoña Aran ◽  
Laia Miquel-Serra ◽  
Yolanda Muñoz ◽  
Elisabet Ars ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Alport Syndrome ◽  
Autosomal Recessive ◽  
Induced Pluripotent

scholarly journals Patient-specific iPSC-derived photoreceptor precursor cells as a means to investigate retinitis pigmentosa

eLife ◽  
2013 ◽  
Vol 2 ◽  
Author(s):  
Budd A Tucker ◽  
Robert F Mullins ◽  
Luan M Streb ◽  
Kristin Anfinson ◽  
Mari E Eyestone ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Protein Misfolding ◽  
Autosomal Recessive ◽  
Stop Codon ◽  
Premature Stop Codon ◽  
Photoreceptor Cells ◽  
Precursor Cells ◽  
Patient Specific ◽  
Basal Bodies ◽  
Induced Pluripotent

Next-generation and Sanger sequencing were combined to identify disease-causing USH2A mutations in an adult patient with autosomal recessive RP. Induced pluripotent stem cells (iPSCs), generated from the patient’s keratinocytes, were differentiated into multi-layer eyecup-like structures with features of human retinal precursor cells. The inner layer of the eyecups contained photoreceptor precursor cells that expressed photoreceptor markers and exhibited axonemes and basal bodies characteristic of outer segments. Analysis of the USH2A transcripts of these cells revealed that one of the patient’s mutations causes exonification of intron 40, a translation frameshift and a premature stop codon. Western blotting revealed upregulation of GRP78 and GRP94, suggesting that the patient’s other USH2A variant (Arg4192His) causes disease through protein misfolding and ER stress. Transplantation into 4-day-old immunodeficient Crb1−/− mice resulted in the formation of morphologically and immunohistochemically recognizable photoreceptor cells, suggesting that the mutations in this patient act via post-developmental photoreceptor degeneration.

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